Alexa 488 conjugated donkey anti rat igg

The procedures of immunostaining were described in our previous studies (Wang et al., 2017 (link); Li, Zhang, Sun, and Ding, 2013 ; Li, Zhang et al., 2014 (link)). Brain sections were sequentially incubated overnight at 4°C with primary antibodies, then incubated with an Alexa 488-conjugated donkey anti-rat IgG (1:200; 712-548-153, Jackson ImmunoResearch Laboratories, West Grove, PA), a Alexa 488-conjugated donkey-anti-mouse IgG (1:200; Life Technologies, CA, USA), an Alexa 594-conjugated donkey anti-rabbit IgG (1:200; 711-585-152, Jackson ImmunoResearch Laboratories) for 4 hr in the dark at room temperature. The brain sections were subjected to fluorescence detection using a Nikon FN1 epi-fluorescence microscopy equipped with a CoolSNAP-EZ CCD-camera or an Olympus confocal microscope and analyzed by MetaMorph software (Molecular Devices, USA). The primary antibodies include a rabbit anti-GDNF polyclonal antibody (1:200; SC-328, Santa Cruz Biotechnology, Santa Cruz, CA), a rabbit anti-Ki67 (1:600, AB9260, Sigma), a rabbit anti-doublecortin (DCX) (1:1000; Ab18723, Abcam, MA. RRID:AB_732011), a mouse anti-S100β monoclonal antibody (1:500; S2532, Sigma. RRID:AB_477499), a mouse anti-GFAP (1:800; G3893, Sigma. RRID:AB_477010), a rat anti-Brdu (1:400; Ab6326, Abcam. RRID:AB_305426), and a rabbit G6PD (1:400; NB100-236, Novus Biologocals, Centennial, CO. RRID:AB_2107663).